Pure platelet-rich plasma promotes semaphorin-3A expression: a novel insight to ameliorate intervertebral disk degeneration in vitro.

INTRODUCTION: Intervertebral disk degeneration (IVDD) can be effectively treated using platelet-rich plasma (PRP). While the exact process is fully understood, it is believed that using pure PRP (P-PRP) without leukocytes is a better option for preventing IVDD. Semaphorin-3A (Sema3A), an inhibitor of angiogenesis and innervation, is essential for preserving IVDD's homeostasis. Whether PRP prevents IVDD by modifying Sema3A has yet to receive much research. This work aims to clarify how P-PRP affects Sema3A when IVDD develops in vitro. METHODS: Nucleus pulposus cells (NPCs) isolated from 8-week-old male Sprague-Dawley rats were exposed to 10 ng/ml IL-1ß and then treated with P-PRP or leukocyte platelet-rich plasma (L-PRP) in vitro, followed by measuring cell proliferation, apoptosis and microstructures, inflammatory gene and Sema3A expression, as well as anabolic and catabolic protein expression by immunostaining, quantitative real-time polymerase chain reaction (qPCR), western blot, and enzyme-linked immunosorbent assay (ELISA). RESULTS: In comparison with L-PRP, P-PRP had a higher concentration of growth factors but a lower concentration of inflammatory substances. P-PRP increased the proliferation of NPCs, while IL-1 relieved the amount of apoptosis due to its intervention. Anabolic genes, aggrecan, and collagen II had higher expression levels. MMP-3 and ADAMTS-4, two catabolic or inflammatory genes, showed lower expression levels. Sema3A activity was enhanced after P-PRP injection, whereas CD31 and NF200 expression levels were suppressed. CONCLUSIONS: P-PRP enhanced the performance of NPCs in IVDD by modifying the NF-κB signaling pathway and encouraging Sema3A expression, which may offer new therapy options for IVDD. THE TRANSLATIONAL POTENTIAL OF THIS ARTICLE: The findings provide a new therapeutic target for the treatment of IVDD and show a novel light on the probable mechanism of PRP and the function of Sema3A in the progression of IVDD.

[Characteristics, Sources, and Contributions to Ozone Formation of Ambient Volatile Organic Compounds in Huanggang, China].

Based on the offline sampling data of volatile organic compounds (VOCs) and the simultaneous online measurements of conventional gaseous air pollutants and meteorological parameters in urban Huanggang, the volume fractions and component characteristics of VOCs were analyzed. The sources and ozone (O3) formation sensitivity of VOCs during severe ozone pollution episodes were analyzed using the positive matrix factorization (PMF) model and the photochemical box model coupled with master chemical mechanism (PBM-MCM), respectively. The results revealed that the average volume fractions of total volatile organic compounds were (21.57±3.13)×10-9, with higher volume fractions in winter and spring compared to those in summer and autumn. Among these, alkanes (49.9%) and alkenes (16.4%) accounted for the highest proportion. The PMF analysis results showed that fuel combustion (27.8%), vehicle emission (19.9%), solvent use (15.7%), industrial halogenated hydrocarbon emission (12.1%), chemical enterprise emission (10.5%), natural sources (7.8%), and diesel vehicle emission (6.2%) were the main sources of VOC emissions. Anthropogenic VOCs emitted by solvent use, fuel combustion, and chemical enterprises contributed significantly (60.9% in total) to generating O3, which indicates that these three types of anthropogenic sources should be controlled first when it comes to preventing and controlling ozone pollution. Further, the relative incremental reactivity (RIR) and empirical kinetic method approach (EKMA) revealed that O3 formation was in a VOCs-limited regime during the observation period in Huanggang, China. Furthermore, O3 formation was more sensitive to m-xylene, p-xylene, ethylene, 1-butene, and toluene; therefore, reducing these VOCs should be prioritized.

GWAS of Chronic Spontaneous Urticaria Reveals Genetic Overlap with Autoimmune Diseases, Not Atopic Diseases.

Although chronic spontaneous urticaria (CSU) is a common disease, GWASs of CSU are lacking. We aimed to identify susceptibility SNPs by performing a GWAS in Chinese Han adults with CSU. The discovery cohort included 430 CSU cases and 482 healthy controls. The GWAS findings were validated in 800 CSU cases and 900 healthy controls. Genetic, functional enrichment, and bioinformatic analyses of genome-wide significant SNPs were performed to assess the association between CSU and autoimmunity or atopy. Five genome-wide significant SNPs were identified: rs434124/LILRA3, rs61986182/IGHG1/2, rs73075571/TDGF1, rs9378141/HLA-G, and rs3789612/PTPN22. The first four SNPs were in linkage disequilibrium with autoimmune-related diseases‒associated SNPs and were cis-expression quantitative trait loci in immune cells. The five SNPs-annotated genes were significantly enriched in immune processes. Higher polygenic risk scores and allele frequencies of rs3789612∗T, rs9378141∗C, and rs73075571∗G were significantly associated with autoimmune-related CSU phenotypes, including positive antithyroglobulin IgG, positive anti-FcεRIα IgG, total IgE <40 IU/ml, and positive antithyroid peroxidase IgG but not with atopic or allergic sensitized CSU phenotypes. This GWAS of CSU identifies five risk loci and reveals that CSU shares genetic overlap with autoimmune diseases and that genetic factors predisposing to CSU mainly manifest through associations with autoimmune traits.

IgE and IgG Anti-Thyroid Autoantibodies in Chinese Patients With Chronic Spontaneous Urticaria and a Literature Review.

Immunoglobulin (Ig) E and IgG anti-thyroid autoantibodies (AAbs) play important roles in the immunopathogenesis of chronic spontaneous urticaria (CSU). To date, association of IgE and IgG AAbs with Chinese CSU patients has not been fully investigated. We aimed to explore prevalence rates of IgE and IgG AAbs in Chinese CSU patients and their association with clinical and laboratory parameters. Serum IgE and IgG AAbs against thyroid peroxidase (TPO) and thyroglobulin (TG), total IgE (tIgE) and specific IgEs were measured using enzyme-linked immunosorbent assay, chemiluminescence microparticle immunoassay and immunoblotting. Meta-analyses and literature review were conducted. The meta-analyses indicated that CSU cases were 4.98, 6.90 and 6.68 times more likely to have positive anti-TPO IgE, anti-TPO IgG and anti-TG IgG (all P < 0.001) compared with controls, respectively, and revealed a positive correlation between the prevalence rates of anti-TPO IgE and anti-TPO IgG (r = 0.53, P = 0.025). A total of 1,100 Chinese Han adult CSU patients and 1,100 ethnicity-, age- and sex-matched healthy controls were recruited from 15 centers. Prevalence rates of anti-TPO IgE, anti-TPO IgG, anti-TG IgE or anti-TG IgG in the patients were all significantly higher than those in the controls. Significant correlations were observed between prevalence rates of anti-TPO IgE and anti-TPO IgG (r = 0.297, P < 0.001) as well as between those of anti-TG IgE and anti-TG IgG in the patients (r = 0.137, P < 0.001). Patients with anti-TPO IgE or anti-TPO IgG had significantly lower tIgE levels (P < 0.001). Positive anti-TPO IgE, positive anti-TPO IgG and tIgE < 40 IU/mL were independent predictors of antihistamine-refractory cases. In conclusion, the prevalence rates of IgE and IgG AAbs in Chinese CSU patients are significantly elevated and reciprocally correlated. This study verifies the results of previous case-control studies of CSU patients from other populations and ethnicities.

Optimal Operation of an Oscillatory Flow Crystallizer: Coupling Disturbance and Stability.

In the process of industrial crystallization, it is always difficult to balance the secondary nucleation rate and metastable zone width (MSZW). Herein, we report an experimental and numerical study for the cooling crystallization of paracetamol in an oscillatory flow crystallizer (OFC), finding the optimal operating conditions for balancing the secondary nucleation rate and MSZW. The results show that the MSZW decreases with the increase of oscillation Reynolds number (Re o). Compared to the traditional stirring system, the OFC has an MSZW three times larger than that of the stirring system under a similar power density of consumption. With the numerical simulation, the OFC can produce a stable space environment and instantaneous strong disturbance, which is conducive to the crystallization process. Above all, a high Re o is favorable to produce a sufficient nucleation rate, which may inevitably constrict the MSZW to a certain degree. Then, the optimization strategy of the operating parameter (Re o) in the OFC is proposed.

[Methods, perspectives and academic innovation, prospects-a review on Rethinking Chinese Acupuncture: A History of Ideas written by professor ZHANG Shu-jian].

In Rethinking Chinese Acupuncture: A History of Ideas, in view of interpretation method and historical perspective, a part of traditional theory of acupuncture and moxibustion is reinterpreted on its concepts and connotations, which endows a fresh significance. By collecting the academic history of acupuncture and moxibustion, the emerging new techniques of acupuncture and moxibustion at home and abroad are summarized. It is suggested that understanding on acupuncture-moxibustion theory should be innovated and progressed with era for the exploration and the construction of novel system of modern acupuncture-moxibustion theory.

The Impact of microRNA Regulation on Immune Recovery in HIV-1-Infected Patients Treated during Acute Infection: A Pilot Study.

microRNAs (miRNAs) are small noncoding RNAs involved in a large range of cellular activities and can be used as biomarkers and indicators for diagnosis. We investigated the alterations in miRNA profiles in immune reconstituted vs. nonimmune reconstituted HIV-1-infected individuals to assess the association between miRNAs and the occurrence of immunological nonresponses, with the aim of searching for miRNA-based biomarkers for these HIV-1-infected individuals. Thirteen immunological responders (IRs) and 12 immunological nonresponders (INRs) were recruited, and RNA was collected from the plasma samples of the 25 HIV-1-infected individuals at both baseline and after 24 months of maintaining virological suppression (VS). Next-generation sequencing was used to detect miRNAs and evaluate the expression differences in miRNAs between IR and INR patients and between baseline and after 24 months of maintaining VS. Samples from 13 IRs and 11 INRs were successfully sequenced. The horizontal comparison of differentially expressed miRNAs between the groups and the longitudinal comparison of differentially expressed miRNAs between baseline and after 24 months of maintaining VS showed that a large proportion of miRNAs in INRs are downregulated compared to the levels in IRs. We also found that the miRNA let-7d-5p was downregulated in 9 INRs but only in 2 IRs by more than 2-fold. The difference was significant. In summary, these results demonstrate for the first time that a large proportion of miRNAs are downregulated in INRs compared with IRs, and the miRNA let-7d-5p is a potential biomarker for INRs.

Single Agent and Synergistic Activity of Maritoclax with ABT-263 in Nasopharyngeal Carcinoma (NPC) Cell Lines.

The BCL-2 anti-apoptotic proteins are over-expressed in many cancers and hence are attractive therapeutic targets. In this study, we tested the sensitivity of two Nasopharyngeal Carcinoma (NPC) cell lines HK1 and C666-1 to Maritoclax, which is reported to repress anti-apoptotic protein MCL-1 and BH3 mimetic ABT-263, which selectively inhibits anti-apoptotic proteins BCL-2, BCL-XL and BCL-w. We investigated the sensitisation of the NPC cell lines to these drugs using the SYBR Green I assay and 3D NPC spheroids. We report that Maritoclax repressed anti-apoptotic proteins MCL-1, BCL-2, and BCL-XL in a dose- and time-dependent manner and displayed a single agent activity in inhibiting cell proliferation of the NPC cell lines. Moreover, combination of Maritoclax and ABT-263 exhibited synergistic antiproliferative effect in the HK1 cells. Similar results were obtained in the 3D spheroids generated from the HK1 cells. More notably, 3D HK1 spheroids either treated with single agent Maritoclax or combination with ABT-263, over 10 days, did not develop resistance to the treatment rapidly. Collectively, the findings illustrate that Maritoclax as a single agent or combination with BH3 mimetics could be potentially useful as treatment strategies for the management of NPC.

Clinical characteristics, diagnosis, and treatment of COVID-19: A case report.

BACKGROUND: Since December 2019, many cases of pneumonia caused by novel coronavirus have been discovered in Wuhan, China, and such cases have spread nationwide quickly. At present, coronavirus disease 2019 (COVID-19) is a worldwide pandemic. What are the clinical features of this disease? What is the clinical diagnosis and how should such patients be treated? As a clinician, mastery of the clinical characteristics, basic diagnosis, and treatment methods of COVID-19 are required to provide help to patients. CASE SUMMARY: A 42-year-old male patient with a cough lasting 6 d without obvious cause, as well as fever and fatigue for 1 d, was admitted to Hankou Hospital on January 22, 2020 and transferred to Huoshenshan Hospital on February 4. The main clinical symptoms were dry cough, fatigue, and fever. He was diagnosed with COVID-19. From the 4th d of admission, the patient's condition gradually worsened, with increased respiratory rate and body temperature. Peripheral blood lymphocytes decreased progressively. On the 8th d of admission, the patient's highest temperature was 40.7 °C, and oxygen saturation was 83% despite high-flow oxygen inhalation. Chest computed tomography results showed that the virus progressed rapidly. The number of lesions significantly increased with expanded scope and increased density. The distribution of lesions advanced from peripheral to central. In addition to nasal catheter oxygen inhalation and symptomatic support, antiviral drugs were used throughout the treatment. On January 22, oseltamivir phosphate capsules were given orally (75 mg, twice daily) for 6 d. On January 24, three tablets of lopinavir and ritonavir were added orally (twice daily). After 6 d, this was changed to 0.2 g (two tablets) arbidol, taken orally (three times daily) for 5 d. During the severe stage, methylprednisolone was given (40 mg) once every 12 h, immunoglobulin (20 g) was administered by intravenous drip infusion once daily, and thymosin (1.6 mg) was injected subcutaneously once daily combined with immunotherapy. On February 2, symptoms decreased, various indicators improved, and pulmonary inflammation was obviously reduced. Throat swabs on February 4 and 9 were negative for novel coronavirus nucleic acid. After 19 d in the hospital, the patient was successfully treated and discharged. CONCLUSION: COVID-19 in young adults can be successfully treated with active treatment. We report a typical case of COVID-19, analyze its clinical characteristics, summarize its clinical diagnosis and treatment experience, and provide a reference for clinical colleagues.

PBF, a Proto-oncogene in Esophageal Carcinoma.

Emerging evidence shows that the pituitary tumour-transforming gene (PTTG)-binding factor (PBF) functions as a proto-oncogene in some tumors. However, the precise functions of PBF in tumorigenesis and its action mechanisms remain largely unknown. Here for the first time we demonstrated that PBF was associated with a tumor-related cell phenotype in esophageal carcinoma (ESCA) and identified the involved signaling pathways. PBF was up-regulated in ESCA tissues (Data from GEPIA) and cells. Then we down-regulated PBF in ESCA cell lines, Eca-109 and TE-1, by using RNAi technology. Cell function analysis suggested that down-regulation of PBF could inhibit tumor-related cell phenotypes, including proliferation, motility, apoptosis and cell cycle, in Eca-109 and TE-1 cells. Mechanism investigation suggested that apoptosis induced by PBF knockdown may be mediated by the activation of the mitochondrial apoptosis pathway and cell cycle arrest. AKT/mTOR and Wnt3a/ß-catenin, key pathways in regulating tumor proliferation and metastasis, were found to be inactivated by the down-regulation of PBF in ESCA cells. In conclusion, our study demonstrates that PBF functions as a proto-oncogene in ESCA in vitro, which may be mediated through AKT/mTOR and Wnt3a/ß-catenin pathways.

Comparison of the value of superficial mark guided localization and hook-wire guided localization techniques for non-palpable breast microcalcifications: A retrospective clinical research.

AIM: The present study compares the effect and accuracy of the superficial mark guided localization (SGL) and hook-wire guided localization (WGL) techniques for non-palpable breast microcalcifications. METHODS: This retrospective study was conducted to compare SGL and WGL techniques. These techniques were performed on 51 patients with non-palpable breast microcalcifications from January 2015 to May 2016. RESULTS: Among these 51 patients, 25 (49.01%) patients were subjected to WGL and 26 patients (50.99%) were subjected to SGL. The SGL technique had a higher rate of malignant cancer detection (WGL = 12.0% and SGL = 23.0%). Furthermore, no significant differences were found with regard to average age, the rate of a second excision and the diameter of the excised tissue. Moreover, no complications were observed in the SGL group, while four (16.0%) patients in the WGL group experienced problems. CONCLUSION: The SGL technique is as accurate as the WGL technique. Furthermore, the procedure has advantages of being less expensive and causing less complications.

Development of a TaqMan probe-based quantitative reverse transcription PCR assay for detection of Getah virus RNA.

Getah virus (GETV), a mosquito-borne virus that mainly infects horses and pigs, has emerged and spread in China. We developed a highly specific and reproducible TaqMan probe-based quantitative reverse transcription PCR (RT-qPCR) assay targeting the non-structural protein 1 of GETV, whose detection limit is 25.5 copies/µL, which is 100-fold higher than that of conventional RT-PCR. RT-qPCR was used to detect GETV RNA in mosquito and animal clinical samples, showing that the accuracy of RT-qPCR was higher than that of conventional RT-PCR. The newly developed RT-qPCR assay may be a useful alternative tool for rapid, simple and specific diagnosis of GETV infection.

WW domain-containing E3 ubiquitin protein ligase 1 depletion evokes antitumor effect in cutaneous squamous cell carcinoma by inhibiting signal transducer and activator of transcription 3 signaling pathway.

Objectives WW domain-containing E3 ubiquitin protein ligase 1 (WWP1) has been implicated in tumor progression. We aimed to investigate the role of WWP1 in cutaneous squamous cell carcinoma (CSCC). Methods WWP1 gene and protein levels were detected using semi-quantitative reverse transcription-polymerase chain reaction, immunohistochemistry and western blotting. The effects of WWP1 on cell cycle, apoptosis, cell migration and invasion were examined by flow cytometry, wound healing and Transwell assays, respectively. The antitumor efficacy of WWP1 small interfering RNA was determined in CSCC tumor xenografts in mice. Results WWP1 expression was significantly higher in CSCC tissues and cells than in normal skin and cells, respectively. WWP1 expression was significantly associated with histological grade, invasion depth and lymph node metastasis in patients with CSCC. High expression predicted metastatic potential and an unfavorable prognosis. WWP1 downregulation suppressed tumor growth in vitro and in vivo, reduced cell migration and invasion, arrested the cell cycle in G0/G1 and induced apoptosis in A431 cells. WWP1 depletion also decreased phosphorylated signal transducer and activator of transcription 3 (STAT3), matrix metalloproteinase-2, cyclin D1 and Bcl-2, but did not affect total STAT3. Conclusions WWP1 is a potential target for the diagnosis, prognosis and therapy of patients with CSCC.

Combined pretreatment serum CA19-9 and neutrophil-to-lymphocyte ratio as a potential prognostic factor in metastatic pancreatic cancer patients.

The aim of this study was to explore the role of combined pretreatment serum carbohydrate antigen 19-9 (CA19-9) and neutrophil-to-lymphocyte ratio (NLR) as potential prognostic factors in metastatic pancreatic cancer patients.We investigated pretreatment serum CA19-9 and NLR in 59 metastatic pancreatic cancer patients, determined the patients' thresholds by receiver operating characteristic curve analysis, and assessed their prognostic values by Kaplan-Meier curve and Cox regression models.Results of multivariate analysis showed high CA19-9, high NLR, and high score (the scoring system of CA19-9 and NLR) were significantly correlated with overall survival. Area under the curve of the scoring system was higher than that of CA19-9 or NLR.Combined pretreatment serum CA19-9 and NLR is a better prognostic biomarker of metastatic pancreatic cancer patients than CA19-9 or NLR alone.

Potential Application of MicroRNA Profiling to the Diagnosis and Prognosis of HIV-1 Infection.

MicroRNAs (miRNAs) were first identified in Caenorhabditis briggsae and later recognized as playing pivotal roles in a vast range of cellular activities. It has been shown that miRNAs are an important mechanism not only for host defense against virus but also for the establishment of viral infection. During human immunodeficiency virus type 1 (HIV-1) infection, host miRNA profiles are altered either as a host response against the virus or alternatively as a mechanism for the virus to facilitate viral replication and infection or to maintain latency. The altered miRNA profiles can be detected and quantified by various advanced assays, and potentially serve as more sensitive, accurate and cost-efficient biomarkers for HIV-1 diagnosis and disease progression than those detected by currently available standard clinical assays. Such new biomarkers are critical for optimizing treatment regimens. In this review, we focus on the potential application of miRNA profiling to the diagnosis of HIV-1 infection and the monitoring of disease progression.

Prognosis of Hippocampal Function after Sub-lethal Irradiation Brain Injury in Patients with Nasopharyngeal Carcinoma.

This work emphasizes the value of assessing hippocampal function by making a timely MRI-based prognosis following a minor dose of hippocampal irradiation after nasopharyngeal carcinomas (NPC) radiotherapy. A quasi-experiment with case-control design and functional assessments (e.g., neuroimaging analysis with fMRI) was conducted to assess hippocampal function after radiotherapy. We delivered 70 Gy of irradiation to nasopharyngeal carcinomas by 6MV helical radiotherapy and collected data from twenty NPC patients and 24 healthy age-matched subjects. Inevitably, hippocampi also received an average dose of 6.89 Gy (range, 2.0-14 Gy). Seed-based functional connectivity of the hippocampus was applied to estimate the cognitive alteration by time before, one month, and four months after irradiation. Afterward, longitudinal-and-cross-sessional statistical inference was determined with time-dependent measurement analysis of variance (ANOVA) with controlled covariance. Over time, there were longitudinal changes in the functional connectivity of hippocampal-related cortices, including the right middle frontal lobe, left superior temporal lobe, and left postcentral gyrus. The findings indicate the presence of functional plasticity, demonstrating how minor irradiation affects functional performance during the early delayed phase of irradiation-induced brain injury.

Modeling nasopharyngeal carcinoma in three dimensions.

Nasopharyngeal carcinoma (NPC) is a type of cancer endemic in Asia, including Malaysia, Southern China, Hong Kong and Taiwan. Treatment resistance, particularly in recurring cases, remains a challenge. Thus, studies to develop novel therapeutic agents are important. Potential therapeutic compounds may be effectively examined using two-dimensional (2D) cell culture models, three-dimensional (3D) spheroid models or in vivo animal models. The majority of drug assessments for cancers, including for NPC, are currently performed with 2D cell culture models. This model offers economical and high-throughput screening advantages. However, 2D cell culture models cannot recapitulate the architecture and the microenvironment of a tumor. In vivo models may recapitulate certain architectural and microenvironmental conditions of a tumor, however, these are not feasible for the screening of large numbers of compounds. By contrast, 3D spheroid models may be able to recapitulate a physiological microenvironment not observed in 2D cell culture models, in addition to avoiding the impediments of in vivo animal models. Thus, the 3D spheroid model offers a more representative model for the study of NPC growth, invasion and drug response, which may be cost-effective without forgoing quality.